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Found 8 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
| This kit uses a green fluorescent probe that reacts with autophagic vacuoles and is detectable by flow cytometry |
Use Cyto-ID green dye (1 μl/4 ml assay buffer) for 30 minutes in dark at RT. |
Incase of Low CYTO-ID Green dye staining in all treatments, increase the reagent concentration (500X dilution of the dye is recommended) and the incubation time.
Incase of High CYTO-ID Green staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experiment.
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| Upstream tips |
| This kit uses a green fluorescent probe that reacts with autophagic vacuoles and is detectable by flow cytometry |
| Protocol tips |
| Use Cyto-ID green dye (1 μl/4 ml assay buffer) for 30 minutes in dark at RT. |
| Downstream tips |
Incase of Low CYTO-ID Green dye staining in all treatments, increase the reagent concentration (500X dilution of the dye is recommended) and the incubation time.
Incase of High CYTO-ID Green staining, it could be that the cell culture medium was depleted of nutrients and so change media 4-8 hours before the experiment.
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| Upstream tips |
Protocol tips |
Downstream tips |
| Intercalates between DNA/RNA and used to analyze mitochondria and Lysosomal content by flow cytometry. |
Stain cells with acridine orange (1 μg/ml) for 15 mins at 37°C |
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| Upstream tips |
| Intercalates between DNA/RNA and used to analyze mitochondria and Lysosomal content by flow cytometry. |
| Protocol tips |
| Stain cells with acridine orange (1 μg/ml) for 15 mins at 37°C |
| Upstream tips |
Protocol tips |
Downstream tips |
| Highly potent inhibitor for mTOR and DNA-PK while inhibits PI3-K only at much higher concentration |
Culture the cells in DMEM medium containing 250nM Torin1 |
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| Upstream tips |
| Highly potent inhibitor for mTOR and DNA-PK while inhibits PI3-K only at much higher concentration |
| Protocol tips |
| Culture the cells in DMEM medium containing 250nM Torin1 |
| Upstream tips |
Protocol tips |
Downstream tips |
| FDA approved Cox-2 inhibitor. It is used to induce autophagy and apoptosis in cells. |
Treat cells with 30 μM of celecoxib for 72 hours |
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| Upstream tips |
| FDA approved Cox-2 inhibitor. It is used to induce autophagy and apoptosis in cells. |
| Protocol tips |
| Treat cells with 30 μM of celecoxib for 72 hours |
| Upstream tips |
Protocol tips |
Downstream tips |
| It is an autophagosome inhibitor. |
Use 30mg/kg/d for in vivo treatment in mice and 2nM for in vitro application in cell line |
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| Upstream tips |
| It is an autophagosome inhibitor. |
| Protocol tips |
| Use 30mg/kg/d for in vivo treatment in mice and 2nM for in vitro application in cell line |
| Upstream tips |
Protocol tips |
Downstream tips |
| It is an autophagosome inhibitor. |
Use 30mg/kg/d for in vivo treatment in mice and 2nM for in vitro application in cell line |
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| Upstream tips |
| It is an autophagosome inhibitor. |
| Protocol tips |
| Use 30mg/kg/d for in vivo treatment in mice and 2nM for in vitro application in cell line |
| Upstream tips |
Protocol tips |
Downstream tips |
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Incubate with 50 μM of MDC in PBS 1x at 37 °C for 15 min |
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| Protocol tips |
| Incubate with 50 μM of MDC in PBS 1x at 37 °C for 15 min |
| Upstream tips |
Protocol tips |
Downstream tips |
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For Co-IP, Use non-denaturating lysis solution to lyse the cells and incubate the cell lysate for 3 hrs at 4°C with Anti LC3 Antibody.Protein A/G then can be incubated overnight. |
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| Protocol tips |
| For Co-IP, Use non-denaturating lysis solution to lyse the cells and incubate the cell lysate for 3 hrs at 4°C with Anti LC3 Antibody.Protein A/G then can be incubated overnight. |
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